Monday, June 6, 2011

Nextera technology featured at ASM 111th General Meeting

Less is more--Evaluation of a low-input, transposase-mediated protocol for rapid generation of high-throughput sequence libraries

Epicentre’s Nextera™ library preparation kits have taken the metagenomics field by storm. Researchers are taking advantage of the rapid, simple Nextera library preparation to determine species, as well as specific enzyme activities, present in a given environmental sample. In a poster at the 111th General Meeting of the American Society for Microbiology (ASM) in New Orleans, Marine et al. used the Nextera DNA Sample Prep Kit (Roche 454-Compatible) to dissect a mock sample in a cultivation-independent bacteriophage preparation. The library, consisting of five mycobacteriophages and four cyanophages, had a relatively broad range of GC content (35.3%-64.7%) and read abundance (28.1%-0.3%).

The results showed that the Nextera library preparation did not compromise the relative abundance of each of the phages in the preparation, and scarce members of the population remained detectable in the presence of more populous library members. The assembled genomes of four of the phages from the Nextera-generated sequence data covered >99% of the genomes at near-perfect identity. GC content was found to affect the coverage of the genomes, though the researchers attributed part of the GC bias to amplification bias in the emulsion PCR prior to sequencing.

The researchers concluded that the Nextera technology is a good alternative to older library preparation methods, and generated quality sequence data from a variety of origins from small quantities (50 ng) of input DNA.

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